华南理工大学学报(自然科学版) ›› 2010, Vol. 38 ›› Issue (9): 127-131.doi: 10.3969/j.issn.1000-565X.2010.09.024

• 生物工程 • 上一篇    下一篇

酶法合成柚皮苷棕榈酸酯的色谱分离及结构鉴定

黄建蓉 李冰 张霞 李琳   

  1. 华南理工大学 轻工与食品学院, 广东 广州 510640
  • 收稿日期:2010-01-04 修回日期:2010-02-12 出版日期:2010-09-25 发布日期:2010-09-25
  • 通信作者: 李琳(1962-),男,教授,博士生导师,主要从事糖类物质及其药物制备与生物利用研究.E—mail:felinli@scut.edu.cn. E-mail:joanjr@126.com
  • 作者简介:黄建蓉(1975-),女,博士生,广东药学院高级工程师,主要从事糖类物质及其药物制备与生物利用研究.
  • 基金资助:

    国家“十一五”科技支撑计划项目(2006BAD27B04); 国家自然科学基金资助项目(20976061); 广东省自然科学基金资助项目(9151063201000066)

Chromatography Separation and Structure Identification of Enzymatically-Synthesized Naringin Palmitate

Huang Jian-rong  Li Bing  Zhang Xia  Li Lin   

  1. School of Light Industry and Food Sciences,South China University of Technology,Guangzhou 510640,Guangdong,China
  • Received:2010-01-04 Revised:2010-02-12 Online:2010-09-25 Published:2010-09-25
  • Contact: 李琳(1962-),男,教授,博士生导师,主要从事糖类物质及其药物制备与生物利用研究.E—mail:felinli@scut.edu.cn. E-mail:joanjr@126.com
  • About author:黄建蓉(1975-),女,博士生,广东药学院高级工程师,主要从事糖类物质及其药物制备与生物利用研究.
  • Supported by:

    国家“十一五”科技支撑计划项目(2006BAD27B04); 国家自然科学基金资助项目(20976061); 广东省自然科学基金资助项目(9151063201000066)

摘要: 采用制备高效液相色谱对叔戊醇中固定化脂肪酶Novozym 435催化合成的柚皮苷棕榈酸酯进行分离纯化,利用红外光谱、质谱、核磁共振碳谱对产物进行分析,并对酶催化的区域选择性反应机理进行探讨.确定了适宜的色谱条件:C18色谱柱(21.5 mm×250.0mm,10μm),流动相为甲醇,流速为20 mL/min,进样质量浓度为30 mg/mL,进样体积为3mL,此条件下分离得到的产物纯度〉99%;产物为柚皮苷棕榈酸单酯,酰基化选择性地发生在柚皮苷的葡萄糖基的6″-羟基位上.

关键词: 柚皮苷棕榈酸酯, 脂肪酶, 高效液相色谱, 结构, 区域选择性

Abstract:

First,preparative high-pressure liquid chromatography was used to isolate and purify the naringin palmitate enzymatically synthesized by immobilized lipase Novozym 435 in the t-amyl alcohol system.Next,IR spectrum,mass spectrum and 13C NMR were used to identify the purified product,and the mechanism of regioselectivity of the reaction was discussed.Then,the feasible operation conditions were determined as follows: a C18 column(21.5mm×250.00mm,10μm) with methanol as the mobile phase at a flowrate of 20mL/min,an injection volume of 3mL with the concentration of 30mg/mL.In these conditions,the product with a purity of more than 99% was obtained.Finally,the purified product was identified as monoester of naringin and palmitic acid,and the regioselective acylation was found preferring the 6″-hydroxyl group of naringin glucose.

Key words: naringin palmitate, lipase, high-pressure liquid chromatography, structure, regioselectivity